Alteration of the curved helical structure located in the upstream region of the β-lactamase promoter of plasmid pUC19 and its effect on transcription

Takashi Ohyama, Masahiko Nagumo, Yoshiko Hirota, Sadatoshi Sakuma

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The region preceding the β-lactamase promoter of Escherichia coli plasmid pUC19 has a curved DNA (bent DNA) structure. The center of the curvature was revealed to exist around nucleotide position 2580 of the plasmid, which is just beside RNA polymerase binding region. It was indicated that the identified region is curved even at 60°C. The gross geometry of the curvature was altered by inserting synthetic doubleffcfanded oligonucleotides between positions 2585 and 2586. Effect of the alteration on strength of the promoter was not detected in vitro. However, in vivo analyses showed that the promoter strength is apparently dependent, in part, on the gross geometry of the curvature. Insertions of 4 and 16 bp, both of which altered the gross geometry of the curvature greatly, caused considerable reductions of in vivo level of β-lactamase mRNA. In vivo, overall three-dimensional structure of the region covering the promoter and the curvature seems to play some significant role in transcription of the gene.

Original languageEnglish
Pages (from-to)1617-1622
Number of pages6
JournalNucleic Acids Research
Volume20
Issue number7
Publication statusPublished - 1992
Externally publishedYes

Fingerprint

Transcription
Genetic Promoter Regions
Promoter
Plasmids
Curvature
Gross
Geometry
DNA
DNA-Directed RNA Polymerases
Oligonucleotides
Nucleotides
Escherichia coli
RNA
Messenger RNA
Genes
Escherichia Coli
Insertion
Covering
Gene
Three-dimensional

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Alteration of the curved helical structure located in the upstream region of the β-lactamase promoter of plasmid pUC19 and its effect on transcription. / Ohyama, Takashi; Nagumo, Masahiko; Hirota, Yoshiko; Sakuma, Sadatoshi.

In: Nucleic Acids Research, Vol. 20, No. 7, 1992, p. 1617-1622.

Research output: Contribution to journalArticle

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